Podocin

Manufactured by Proteintech

Sourced in China

Podocin is a laboratory product manufactured by Proteintech. It is a protein essential for the proper functioning of the glomerular basement membrane in the kidney. Podocin plays a critical role in maintaining the structural integrity and selective permeability of the kidney filtration barrier.

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Lab products found in correlation

Nephrin, by Thermo Fisher Scientific (2 mentions) Cd2ap, by Thermo Fisher Scientific (2 mentions) Tnf α, by Wuhan Servicebio Technology (1 mentions) Interleukin 6 (il 6), by Wuhan Servicebio Technology (1 mentions) Il 1β, by Wuhan Servicebio Technology (1 mentions) Tunel kit, by Wuhan Servicebio Technology (1 mentions) Fv3000, by Olympus (1 mentions) Cleaved caspase 3, by Wuhan Servicebio Technology (1 mentions) Beclin1, by Wuhan Servicebio Technology (1 mentions) Bcl 2, by Abcam (1 mentions) Bcl2 associated x (bax), by Abcam (1 mentions) Vectastain dab kit, by Vector Laboratories (1 mentions) Desmin, by Abcam (1 mentions) Sirt1, by Abcam (1 mentions) Enhanced chemi luminescence (ecl), by Tiangen Biotech (1 mentions) Foxo1, by Cell Signaling Technology (1 mentions) Beclin 1, by Proteintech (1 mentions) Ripa lysis buffer, by Applygen (1 mentions) Protease inhibitor cocktail, by CWBIO (1 mentions) Anti fkn, by Abcam (1 mentions)

6 protocols using podocin

Immunofluorescence Analysis of Kidney Injury

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TUNEL kits (Servicebio, G1501) were used to detect apoptotic cells in paraformaldehyde-fixed kidney tissues, and the appropriate experimental protocols were applied. For immunofluorescence in tissues, paraffin sections were blocked with 10% donkey serum and incubated overnight at 4 °C with primary antibodies nephrin (1:100, Invitrogen, PA5-106921), podocin(1:100, Proteintech, 20384-1-AP), cd2ap(1:100, Invitrogen, PA5-51894), TNF-α(1:500, Servicebio, GB11188), IL-6(1:500, Servicebio, GB11117), IL-1β(1:300, Servicebio, GB11113), and LC3(1:500, Proteintech, 14600-1-AP), followed by secondary antibodies. Finally, the nuclei were restained with DAPI and observed under a confocal microscope (Olympus, FV3000).
For immunofluorescence in cells, each group of cells were given the corresponding treatment and sequentially subjected to cell rupture, serum closure, incubated with the corresponding primary antibodies nephrin (1:100, Invitrogen, PA5-106921), podocin (1:100, Proteintech, 20384-1-AP), cd2ap (1:100, Invitrogen, PA5-51894), and LC3(1:500, Proteintech, 14600-1-AP) overnight at 4 °C and secondary antibodies. Finally, after DAPI restaining nuclei, the cells were observed under confocal microscopy.

Ji B., Liu J., Yin Y., Xu H., Shen Q, & Yu J. (2023). Minnelide combined with anti-ANGPTL3-FLD monoclonal antibody completely protects mice with adriamycin nephropathy by promoting autophagy and inhibiting apoptosis. Cell Death & Disease, 14(9), 601.

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Podocyte Marker Evaluation in Rat Kidney Tissue

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The frozen kidney tissue of rats was cut into 5 μm thick sections for indirect immunofluorescence staining of podocyte markers podocin and nephrin. Rabbit polyclonal antibodies against podocin (1:200 dilution, Proteintech) and nephrin (1:200 dilution, Abcam) were used as primary antibodies. Cy3-labeled goat anti-rabbit immunoglobulin (Servicebio) was used as a secondary antibody.
Cells cultured on glass slides were washed twice with cold phosphate-buffered saline (PBS) and fixed in 4% paraformaldehyde for 20 min. After washing thrice with phosphate buffered saline (PBS), the cells were treated with 0.1% Triton X-100 for 5 min, blocked with 3% fetal bovine serum (FBS) in PBS for 30 min at room temperature, incubated with the specific primary antibody, podocin (1:200 dilution, Proteintech), and then stained with Cy3-labeled secondary antibody. Semi-quantitative analysis of specific protein expression was performed using fluorescence microscopy, and the mean gray value, representing the mean fluorescence intensity, was analyzed using ImageJ.

Liu H., Wang J., Yue G, & Xu J. (2024). Placenta-derived mesenchymal stem cells protect against diabetic kidney disease by upregulating autophagy-mediated SIRT1/FOXO1 pathway. Renal Failure, 46(1), 2303396.

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Kidney Podocyte Protein Expression Analysis

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Kidney tissues and podocytes were first lysed with RIPA, followed by electrophoresis, membrane transfer, blocking, probing with primary antibodies nephrin (1:500, Invitrogen, PA5-106921), podocin (1:500, Proteintech, 20384-1-AP), cd2ap (1:500, Invitrogen, PA5-51894), Bax (1:500, Abcam, ab216494), Bcl-2 (1:500, Abcam, ab196495), Cleaved-caspase3 (1:500, Servicebio, GB11532), p-mTOR (1:1000,CST,5536 T), mTOR (1:1000,CST,2983 T), Beclin1(1:500, Servicebio, GB112053), p62(1:500, Servicebio, GB11239-1), and LC3(1:1000, Proteintech, 14600-1-AP) overnight at 4 °C, and incubation with secondary antibodies for 1 h at room temperature. Finally, the immunoreactive bands were developed by chemiluminescence and analyzed using Image J software.

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Immunohistochemical Analysis of Glomerular Proteins

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Immunohistochemistry was performed on paraffin sections using a high pressure-based antigen retrieval technique. Hydrogen peroxide (3%) and normal goat serum was used to block endogenous peroxidase activity and nonspecific binding. Primary antibodies used included rabbit anti-nephrin (Abnova, Taipei, Taiwan), podocin (Proteintech, Wuhan, China), and desmin (Abcam, Cambridge, UK). After incubation with primary antibodies at 4 °C overnight, renal sections were reacted with a biotinylated secondary antibody and horseradish peroxidase-streptavidin working buffer (SV-0002 Histostain Plus Kits, Boster Bioengineering, Wuhan, China) at room temperature for 45 min, respectively. A Vecta-stain DAB Kit (Vector Labs, Burlingame, CA) was then used to visualize peroxidase activity. Finally, each slide was counterstained with hematoxylin. The results were scored by semiquantitative immunohistochemical analysis as follows: no staining (−), weak staining (+), moderate staining (++) and strong staining (+++).^{22 (link)} We evaluated staining of 20 randomly selected glomeruli using a 40× objective. Scores for each rat are expressed as the mean value of all scores obtained for that animal.

Gao F., He X., Liang S., Liu S., Liu H., He Q., Chen L., Jiang H, & Zhang Y. (2018). Quercetin ameliorates podocyte injury via inhibition of oxidative stress and the TGF-β1/Smad pathway in DN rats. RSC Advances, 8(62), 35413-35421.

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Protein Extraction and Western Blot Analysis

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Total protein was extracted from MPC5 cells using RIPA lysis buffer (Applygen, Beijing, China), and protein concentration was determined using a protein detection kit (TransGen Biotech, Beijing, China). The primary antibodies used in this study were podocin (1:500; Proteintech); B ECLin-1 (1:1000; Proteintech), LC3 (1:500; Proteintech), FOXO1 (1:1000; Cell Signaling Technology), and SIRT1 (1:1000; Abcam). ECL (Tiangen Biochemical Technology, Beijing, China) was used to detect immunoreactive bands after incubation with the corresponding horseradish peroxidase-conjugated secondary antibodies.

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Western Blot Analysis of Kidney Proteins

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After treatment of cells or kidney tissues for 24 h, total proteins were extracted from cells with RIPA buffer (Beyotime Biotechnology, Shanghai, China) containing a protease inhibitor cocktail (Cwbiotech, Beijing, China) (1:99). Protein samples were quantified, loaded, and separated by 8% sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to a PVDF membrane (GE Healthcare, Freiburg, Germany). After transfer, 5% BSA (in TBST buffer) was used to block the membranes at room temperature for 1 h. Then, the membranes were pre-incubated with different primary antibodies including anti-nephrin, anti-FKN (Abcam, Hongkong, China), podocin, CD2AP, SYNPO, β-catenin (Proteintech, Hubei, china), wnt-4, cyclin-D1 and c-myc (Santa Cruz Biotechology, California, USA), and GAPDH (Danvers, MA, USA) antibodies at 4 °C overnight. After washing with TBST three times, membranes were incubated with secondary antibody. For visualization of detected proteins, immunoblots were analyzed using an enhanced chemiluminescence (ECL, Millipore, Billerica, MA, USA) western blot detection kit and the peroxidase luminescence intensity was measured using the Universal Hood II Molecular Imager GEL System (Bio-Rad, USA).

Senouthai S., Wang J., Fu D, & You Y. (2019). Fractalkine is Involved in Lipopolysaccharide-Induced Podocyte Injury through the Wnt/β-Catenin Pathway in an Acute Kidney Injury Mouse Model. Inflammation, 42(4), 1287-1300.

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